Recommended strains of rhizobia for Trifolium repens and T. subterraneum, and rhizobia isolated from T. angustifolium and T. pratense root nodules, were used to inoculate seven accessions of T. nigrescens and their own macrosymbiont host plants growing in N-free media. When inoculated with the strain of rhizobia recommended for T. subterraneum, foliage weights of six of the seven T. nigrescens accessions were either the highest, or not significantly less than the highest. This supports the inclusion of T. nigrescens in the T. subterraneum group when grouping species which respond similarly symbiotically. The response of foliage weights of T. subterraneum and T. repens plants to inoculation with the strain of rhizobia isolated from T. angustifolium was similar to that following inoculation with the recommended strain for T. repens, rather than with the recommended strain for T. subterraneum. This result indicates that T. angustifolium should be included in the T. repens group when grouping species which respond similarly in symbiosis.